One aspect of this invention relates to the treatment of chronic viral infections by administration of homeopathic dilutions of growth factors. Chronic viral infections, such as herpes simplex virus, Epstein-Barr virus (EBV), human immunodeficiency virus (HIV), papilloma virus, AIDS, chronic fatigue syndrome, Coxsackie B, hauta virus and hepatitis B virus, affect signal transduction mechanisms with deleterious effects within and between the host's immune and nervous systems. During chronic viral infection, host cell signal transduction and cell cycle regulation are altered, often causing cell injury and cell death.
Viruses lack the necessary biochemical machinery to manufacture proteins and must therefore insert their genetic material into a host cell genome in order to proliferate. Viruses consist of a protein coat and genetic material. RNA viruses additionally contain reverse transcriptase, an enzyme that translates the RNA into a DNA strand before insertion in the host cell genome.
During viral infection, the protein coat binds to the host cell's surface membrane enabling the virus' genetic information to subsequently enter the host cell. Entry occurs via various methods, one of which is attachment to specific membrane receptors, including growth factor receptors. For example, the cell receptors for the Epstein Barr and herpes simplex type 1 viruses have been identified as the third component of the complement receptor and the fibroblast growth factor receptor, respectively. Insertion of viral genetic information into the host cell's genome subverts the cell's normal metabolic and genetic mechanisms in order to prioritize viral gene expression and replication.
Chronic, or long-term, viral infections occur when the virus is able to overcome or effectively disrupt the normal neuronal and immunological defense mechanisms of the host. Several viruses, such as herpes simplex virus, EBV, human herpes 6 virus (HH6V), hepatitis B and HIV are able to cause latent (asymptomatic) infections in specific cell populations. Viral replication subsequently occurs in response to extracellular stimuli (Garcia-Blanco, M. A. and Cullen, B. R. 1991 Science 254:815-820). Infections become persistent when continuous viral replication occurs without substantial disruption of host cell function. Viral infections are terminated only when viral replication is disrupted.
Viral infection erodes feedback communication between the host's immune and nervous systems. For example, synthesis of adrenocorticotrophic hormone (ACTH) by lymphocytes after viral infection disrupts the normal feedback loop between pituitary/hypothalamus secretion of ACTH and the adrenal gland's synthesis of glucocorticoids in response to ACTH signals. Over-expression of ACTH causes increased expression of glucocorticoids which consequentially down-regulates the pituitary and suppresses the activities of T lymphocytes. This constant stress response often leads to extreme fatigue and exhaustion in patients with chronic viral infections. In an immune compromised patient, chronic infection leads to entry of virions into the bloodstream, the lymphatic vessels and/or the nerve pathways resulting in infection of new and distant cell populations.
DNA viral infections are often correlated with chronic or cancerous illnesses. For example, hepatitis B viral infection may be correlated with liver cirrhosis of the time and primary hepatocellular carcinoma 58% of the time compared to 17% in a control group. EBV infection correlates with Hodgkin's disease of the mixed cellularity type 60% of the time. Herpes type viral nucleic acid sequences from herpes simplex 1 and 2, cytomegalovirus and EBV have been found in the cerebrospinal fluid of patients with acute encephalitiso In addition, EBV has been found to induce receptors for human herpes 6 virus (HH6V). HH6V has, in turn, been found to be a cofactor in causing chronic fatigue syndrome and AIDS. The ability of viruses to cause cancer is contained within specific sequences of the viral genome. These sequences, known as oncogenes, have the ability to modulate gene transcription and regulation.
Gene transcription and regulation is modulated under normal conditions by growth factors. Growth factors are cell signalling polypeptides that bind to specific cell membrane receptors and initiate a cascade of intracellular events that affect cell proliferation and differentiation. As stated above, many growth factors bind to the same cell surface receptors as viruses and therefore activate the same metabolic pathways used by viral infected or transformed cells.
Growth factors and viruses use the same transcription sites to regulate cell proliferation. For example, TGF.beta. plays a critical role in the transmission of biological information by acting as an on/off switch that couples cell behavior to the external environment. Within the TGF.beta. promoter lies the proto-oncogene c-fos which codes for key transcription factors located at AP-1 transcription sites. Subversion of c-fos gene expression mediates HIV transcription and replication independent of control sites located at tat and NF.sub.k .beta. (Roebuck, K. A. et al. 1993 J. Clin. Invest. 92:1336-1348). Viral transcription in the human T-cell leukemia virus type 1 (HTLV-1), a virus with many characteristics similar to HIV, is tightly regulated by a Tax transactivator site located at the c-fos AP-1 site within the TGF.beta. promoter (Kim et al. 1990 J. Exp. Med. 172:121-129). When the TGF.beta. promoter is activated so is HTLV-1 Tax.
There are homologies between the gene sequences of growth factors, proto-oncogenes and oncogenes. It is now known that normal non-cancer cells contain proto-oncogenes that are homologous to the oncogenes found in some cancer causing viruses. These sequences have the power to regulate the cell cycle. Growth factors regulate the cell cycle by manipulating proto-oncogenes. Some proto-oncogene sequences are homologous with growth factors or their receptors. For example, the B chain of platelet-derived growth factor (PDGF) is homologous to the proto-oncogene c-sis (Doolittle, R. F., et al. 1983 Science 221:275-77). The receptor for epidermal growth factor (EGF) is homologous to the proto-oncogene c-erb.beta. (Downward, et al. 1984 Nature 307:521-527).
Chronic viral infections can lead to up-regulation of growth factor expression. For example, HIV infection up-regulates expression of tumor necrosis factor alpha (TNF.alpha.) and transforming growth factor beta (TGF.beta.). Over-expression of either of these growth factors disrupts normal transcriptional control of gene expression, leading to suppression of hematopoietic progenitor cells and increased HIV replication. TGF.beta., secreted by HIV-infected lymphocytes, also promotes growth of Kaposi's sarcoma cells, fibroblasts and endothelial cells.
Specific hemopoietic growth factors have been used to treat diseases such as AIDS and cancer. Hemopoietic growth factors are logical immunomodulators to use in chronic viral infections and other diseases for several reasons. First, endogenous growth factors such as granulocyte-monocyte colony stimulating factor (GM-CSF) and macrophage colony stimulating factor (M-CSF) stimulate proliferation of hemopoietic progenitor cells. Second, lymphocytes, macrophages and natural killer cells that normally produce these factors are quantitatively and qualitatively defective after infection by HIV, HH6V or EBV. Third, primates infused with GM-CSF showed low toxicity with some positive but inconsistent rises in platelet number.
However, clinical studies on AIDS using GM-CSF and M-CSF at pharmacological doses (ug/kg/day) have produced mixed results. For example, injections or intravenous administration of GM-CSF at concentrations of 0.5-0.8 ug/kg/day transiently increased leukocyte, neutrophil, eosinophil and monocyte counts in AIDS patients with no significant rise in platelet counts or change in reticulocyte and lymphocyte counts (Miles, S. 1992 AIDS Res. Hum. Retroviruses 8:1073-1080). Subcutaneous injections of 0.25-4.0 ug/kg/day improved leukocyte counts with no improvement in hemoglobin or platelet counts. However, the side effects included increased HIV replication, increased levels of P24 antigen, chills, nausea, myalgia and flu-like symptoms (Poli, G. et al. 1991 J. Exp. Med. 173:589-597; Scadden, D. T. 1990 Hematopoietic Growth Factors in Trans. Med., Wiley-Liss Inc., New York, pp. 163-176). GM-CSF also occasionally caused thrombocytopenia. Granulocyte colony stimulating factor (G-CSF) has been effective in correcting neutropenia with some minor increases in lymphocyte counts. Additionally, hemoglobin and reticulocytes increased in numbers in patients given G-CSF alone or in combination with erythropoietin. However, resumption of treatment with AZT after use of these growth factors lead to severe anemia. Pharmacological doses of growth factors often have harsh side effects.
Homeopathy is founded on the principles of pharmacology except with much higher dilutions (much lower concentrations). Homeopathic medicine dates back to the nineteenth century. One of the basic tenets of homeopathic medicine is that a cure for a disease can be evoked by using a high dilution medicine that resembles but is different from the cause of the disease. Homeopathy is widely accepted as a useful therapeutic throughout Europe, the British Commonwealth countries and India, and has been demonstrated to have characteristic and reproducible effects. A critical review of more than 100 controlled and/or clinical studies of homeopathy determined that patients received positive healing benefits from homeopathy beyond the placebo effect (Kleijnen, J. et al. 1991 Brit. Med. J. 302:316-323).
Many homeopathic medicines are used at concentrations of micrograms (10.sup.-6 M) and nanograms (10.sup.-12 M); however, other homeopathic dilutions exceed Avogadro's number (6.023.times.10.sup.-23). When homeopathic compounds are diluted 1:10, with repeated successions (similar to vortexing) and repetitively diluted by this procedure at least 24 times a potency is achieved (10.sup.-24) that contains essentially no molecules of the original substance. Homeopathic practitioners believe that the potency of a compound increases with increasing dilutions. The standard homeopathic dosage is 10-15 drops of a 10.sup.-12 molar, or 6C, solution administered three times per day. A 6C dilution approximates 1 ng/ml, which is used in cell culture but would be considered a lower than physiological dose when used orally.
Highly dilute homeopathic medicines have been effective in treating some viruses in vivo. Homeopathic dilutions of 1.times.10.sup.-200 to 1.times.10.sup.-1000 of typhoidinum, hydrophobinum, tuberculinum, nux vomica and malandrinum 100% inhibited pock-like lesions on the chorio-allantoic membrane of chicken embryos infected with a DNA virus, chicken embryo virus compared to controls (Singh, L. M. and Gupta, G. 1985 Brit. Homeopathy 74:168-174). Other homeopathic medicines, the same medicines at different homeopathic concentrations or control phosphate buffered solution (PBS), had lesser to no effect.
While the exact mechanism of action of homeopathic medicines is unknown, magnetic image resonance measurements on serial dilutions of substances indicate that the hydroxyl (OH) groups in the solvent of solutions continue to change as dilutions become successively higher (Sacks, A. D. 1983 J. Holistic Med. 5:175-176; Smith, R. and Boericke, G. 1968 J. Am. Inst. Homeopathy 61:197-212; Smith, R. and Boericke, G. 1966 J. Am. Inst. Homeopathy 59:263-279). It is clear that the specific effects of homeopathics are of a non-molecular origin, yet provide potent biological information that is clinically effective. It has been postulated that the effects of highly dilute compounds may use signal transduction pathways that are bioelectromagnetic in nature (Benveniste, J. 1993 Frontier Perspectives 3:13-15).
Bioelectromagnetics underlies biochemical reactions. The science of bioelectromagnetics studies the interactions of electromagnetic fields in living systems (Rubik, R. and Flower, R. G. 1993 Electromagnetic applications in medicine, NIH-OAM panel report to congress; Tenforde, T. S. and Kaune, W. T. 1987 Health Physics 53:585-606). Several studies on the effects of administering electromagnetic signals have been published. For example, Thomas et al. have demonstrated behavioral changes in rats following administration of a cyclotron electromagnetic field which resonates for the signal for unhydrated lithium ions (Thomas J. R. et al. 1986 Bioelectromagnetics 2:349-357). Researchers have also reported inhibition of tumor growth by administration of human interferon alpha (IFN-.alpha.) plus DC current (Sersa, G. and Miklavcic, D. 1990 Molecular Biotherapy 2:165-168).
Electromagnetic signals are believed to exert an effect at the cell membrane. Cell membranes maintain a carefully controlled surface potential that is measurable and reproducible. These potentials are transiently altered by electromagnetic fields or viral attachment, as well as by binding of neurotransmitters, hormones and growth factors. Electrical stimulation of cells temporally changes the cell's membrane potential and evokes consequential changes of RNA, DNA and protein synthesis (Bourguignon, G. J. and Bourguignon, L. Y. 1987 FASEB J. 1:398-402; Rodan, G. A. et al. 1978 Science 190:690-692).
Few effective treatments are available for disorders such as chronic viral infections, cancer and diabetes. Insulin-dependant diabetes, while regulated by insulin still has many systematic complications. Despite more than ten years of aggressive research, both conventional and naturopathic, no definitive treatment exists for HIV infection or acquired immunodeficiency syndrome (AIDS). There thus continues to be a need in the art for effective treatments for chronic viral infections, cancer and diabetes.